Cardiac fibroblasts (CFs) are emerging as major contributors to myocardial fibrosis (MF), a final common pathway of many etiologies of heart disease. Here, we studied the functional relevance of transient receptor potential canonical 3 (TRPC3) channels and nuclear factor of activated T cells c3 (NFATc3) signaling in rodent and human ventricular CFs, and whether their modulation would limit MF.
Results:
A positive feedback loop between TRPC3 and NFATc3 drove a rat ventricular CF fibrotic phenotype. In these cells, polyphenols (extract of grape pomace polyphenol [P.E.]) decreased basal and angiotensin II-mediated Ca2+ entries through a direct modulation of TRPC3 channels and subsequently NFATc3 signaling, abrogating myofibroblast differentiation, fibrosis and inflammation, as well as an oxidative stress-associated phenotype. N(ω)-nitro-l-arginine methyl ester (l-NAME) hypertensive rats developed coronary perivascular, sub-epicardial, and interstitial fibrosis with induction of embryonic epicardial progenitor transcription factors in activated CFs. P.E. treatment reduced ventricular CF activation by modulating the TRPC3-NFATc3 pathway, and it ameliorated echocardiographic parameters, cardiac stress markers, and MF in l-NAME hypertensive rats independently of blood pressure regulation. Further, genetic deletion (TRPC3−/−) and pharmacological channel blockade with N-[4-[3,5-Bis(trifluoromethyl)-1H-pyrazol-1-yl]phenyl]-4-methyl-benzenesulfonamide (Pyr10) blunted ventricular CF activation and MF in l-NAME hypertensive mice. Finally, TRPC3 was present in human ventricular CFs and upregulated in MF, whereas pharmacological modulation of TRPC3-NFATc3 decreased proliferation and collagen secretion.
Innovation and Conclusion:
We demonstrate that TRPC3-NFATc3 signaling is modulated by P.E. and critically regulates ventricular CF phenotype and MF. These findings strongly argue for P.E., through TRPC3 targeting, as potential and interesting therapeutics for MF management.
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References
1.
BraitschCM, KanisicakO, van BerloJH, MolkentinJD, and YutzeyKE. Differential expression of embryonic epicardial progenitor markers and localization of cardiac fibrosis in adult ischemic injury and hypertensive heart disease. J Mol Cell Cardiol, 65: 108–119, 2013.
2.
CassidyA, MukamalKJ, LiuL, FranzM, EliassenAH, and RimmEB. High anthocyanin intake is associated with a reduced risk of myocardial infarction in young and middle-aged women. Circulation, 127: 188–196, 2013.
3.
ChacarS, ItaniT, HajalJ, SalibaY, LoukaN, FaivreJF, MarounR, and FaresN. The impact of long-term intake of phenolic compounds-rich grape pomace on rat gut microbiota. J Food Sci, 83: 246–251, 2018.
4.
ChenJB, TaoR, SunHY, TseHF, LauCP, and LiGR. Multiple Ca2+ signaling pathways regulate intracellular Ca2+ activity in human cardiac fibroblasts. J Cell Physiol, 223: 68–75, 2010.
5.
D'ArchivioM, SantangeloC, ScazzocchioB, VarìR, FilesiC, MasellaR, and GiovanniniC. Modulatory effects of polyphenols on apoptosis induction: relevance for cancer prevention. Int J Mol Sci, 9: 213–228, 2008.
6.
DavisJ, BurrAR, DavisGF, BirnbaumerL, and MolkentinJD. A TRPC6-dependent pathway for myofibroblast transdifferentiation and wound healing in vivo. Dev Cell, 23: 705–715, 2012.
7.
DavisJ and MolkentinJD. Myofibroblasts: trust your heart and let fate decide. J Mol Cell Cardiol, 70: 9–18, 2014.
8.
Del RioD, Rodriguez-MateosA, SpencerJP, TognoliniM, BorgesG, and CrozierA. Dietary (poly)phenolics in human health: structures, bioavailability, and evidence of protective effects against chronic diseases. Antioxid Redox Signal, 18: 1818–1892, 2013.
DobrydnevaY, WilliamsRL, and BlackmorePF. Diethylstilbestrol and other nonsteroidal estrogens: novel class of store-operated calcium channel modulators. J Cardiovasc Pharmacol, 55: 522–530, 2010.
11.
DriesenRB, NagarajuCK, Abi-CharJ, CoenenT, LijnenPJ, FagardRH, SipidoKR, and PetrovVV. Reversible and irreversible differentiation of cardiac fibroblasts. Cardiovasc Res, 101: 411–422, 2014.
12.
EstruchR, RosE, Salas-SalvadoJ, CovasMI, CorellaD, ArosF, Gomez-GraciaE, Ruiz-GutierrezV, FiolM, LapetraJ, Lamuela-RaventosRM, Serra-MajemL, PintoX, BasoraJ, MunozMA, SorliJV, MartinezJA, and Martinez-GonzalezMA. Retraction and republication: primary prevention of cardiovascular disease with a mediterranean diet. N Engl J Med 2013;368: 1279–90. N Engl J Med, 378: 2441–2442, 2018.
13.
FengS, LiH, TaiY, HuangJ, SuY, AbramowitzJ, ZhuMX, BirnbaumerL, and WangY. Canonical transient receptor potential 3 channels regulate mitochondrial calcium uptake. Proc Natl Acad Sci U S A, 110: 11011–11016, 2013.
14.
FujiiT, OnoharaN, MaruyamaY, TanabeS, KobayashiH, FukutomiM, NagamatsuY, NishiharaN, InoueR, SumimotoH, ShibasakiF, NagaoT, NishidaM, and KuroseH. Galpha12/13-mediated production of reactive oxygen species is critical for angiotensin receptor-induced NFAT activation in cardiac fibroblasts. J Biol Chem, 280: 23041–23047, 2005.
15.
GoszczK, DuthieGG, StewartD, LeslieSJ, and MegsonIL. Bioactive polyphenols and cardiovascular disease: chemical antagonists, pharmacological agents or xenobiotics that drive an adaptive response?. Br J Pharmacol, 174: 1209–1225, 2017.
16.
HartmannJ, DragicevicE, AdelsbergerH, HenningHA, SumserM, AbramowitzJ, BlumR, DietrichA, FreichelM, FlockerziV, BirnbaumerL, and KonnerthA. TRPC3 channels are required for synaptic transmission and motor coordination. Neuron, 59: 392–398, 2008.
17.
HaudekSB, XiaY, HuebenerP, LeeJM, CarlsonS, CrawfordJR, PillingD, GomerRH, TrialJ, FrangogiannisNG, and EntmanML. Bone marrow-derived fibroblast precursors mediate ischemic cardiomyopathy in mice. Proc Natl Acad Sci U S A, 103: 18284–18289, 2006.
18.
HeX, LiS, LiuB, SusperreguyS, FormosoK, YaoJ, KangJ, ShiA, BirnbaumerL, and LiaoY. Major contribution of the 3/6/7 class of TRPC channels to myocardial ischemia/reperfusion and cellular hypoxia/reoxygenation injuries. Proc Natl Acad Sci U S A, 114: E4582–E4591, 2017.
19.
HuangJH, HeGW, XueHM, YaoXQ, LiuXC, UnderwoodMJ, and YangQ. TRPC3 channel contributes to nitric oxide release: significance during normoxia and hypoxia-reoxygenation. Cardiovasc Res, 91: 472–482, 2011.
20.
KanisicakO, KhalilH, IveyMJ, KarchJ, MalikenBD, CorrellRN, BrodyMJ, J LinSC, AronowBJ, TallquistMD, and MolkentinJD. Genetic lineage tracing defines myofibroblast origin and function in the injured heart. Nat Commun, 7: 12260, 2016.
21.
KaramaćM. Chelation of Cu(II), Zn(II), and Fe(II) by tannin constituents of selected edible nuts. Int J Mol Sci, 10: 5485–5497, 2009.
22.
KazakovA, HallR, JagodaP, BachelierK, Müller-BestP, SemenovA, LammertF, BöhmM, and LaufsU. Inhibition of endothelial nitric oxide synthase induces and enhances myocardial fibrosis. Cardiovasc Res, 100: 211–221, 2013.
KiyonakaS, KatoK, NishidaM, MioK, NumagaT, SawaguchiY, YoshidaT, WakamoriM, MoriE, NumataT, IshiiM, TakemotoH, OjidaA, WatanabeK, UemuraA, KuroseH, MoriiT, KobayashiT, SatoY, SatoC, HamachiI, and MoriY. Selective and direct inhibition of TRPC3 channels underlies biological activities of a pyrazole compound. Proc Natl Acad Sci U S A, 106: 5400–5405, 2009.
25.
KoppRF, LeechCA, and RoeMW. Resveratrol interferes with Fura-2 intracellular calcium measurements. J Fluoresc, 24: 279–284, 2014.
26.
KramannR, SchneiderRK, DiRoccoDP, MachadoF, FleigS, BondziePA, HendersonJM, EbertBL, and HumphreysBD. Perivascular Gli1+ progenitors are key contributors to injury-induced organ fibrosis. Cell Stem Cell, 16: 51–66, 2015.
27.
KuwaharaK, WangY, McAnallyJ, RichardsonJA, Bassel-DubyR, HillJA, and OlsonEN. TRPC6 fulfills a calcineurin signaling circuit during pathologic cardiac remodeling. J Clin Invest, 116: 3114–3126, 2006.
28.
LighthouseJK and SmallEM. Transcriptional control of cardiac fibroblast plasticity. J Mol Cell Cardiol, 91: 52–60, 2016.
29.
LinCM, ChangH, WangBW, and ShyuKG. Suppressive effect of epigallocatechin-3-O-gallate on endoglin molecular regulation in myocardial fibrosis in vitro and in vivo. J Cell Mol Med, 20: 2045–2055, 2016.
30.
LinZ, MurtazaI, WangK, JiaoJ, GaoJ, and LiPF. miR-23a functions downstream of NFATc3 to regulate cardiac hypertrophy. Proc Natl Acad Sci U S A, 106: 12103–12108, 2009.
NishidaM, OnoharaN, SatoY, SudaR, OgushiM, TanabeS, InoueR, MoriY, and KuroseH. Galpha12/13-mediated up-regulation of TRPC6 negatively regulates endothelin-1-induced cardiac myofibroblast formation and collagen synthesis through nuclear factor of activated T cells activation. J Biol Chem, 282: 23117–23128, 2007.
38.
NoadRL, RooneyC, McCallD, YoungIS, McCanceD, McKinleyMC, WoodsideJV, and McKeownPP. Beneficial effect of a polyphenol-rich diet on cardiovascular risk: a randomised control trial. Heart, 102: 1371–1379, 2016.
Numaga-TomitaT, OdaS, ShimauchiT, NishimuraA, MangmoolS, and NishidaM. TRPC3 channels in cardiac fibrosis. Front Cardiovasc Med, 4: 56, 2017.
41.
OlsonER, NaugleJE, ZhangX, BomserJA, and MeszarosJG. Inhibition of cardiac fibroblast proliferation and myofibroblast differentiation by resveratrol. Am J Physiol Heart Circ Physiol, 288: H1131–H1138, 2005.
42.
OnoharaN, NishidaM, InoueR, KobayashiH, SumimotoH, SatoY, MoriY, NagaoT, and KuroseH.TRPC3 and TRPC6 are essential for angiotensin II-induced cardiac hypertrophy. EMBO J, 25: 5305–5316, 2006.
43.
OrrAL, VargasL, TurkCN, BaatenJE, MatzenJT, DardovVJ, AttleSJ, LiJ, QuackenbushDC, GoncalvesRL, PerevoshchikovaIV, PetrassiHM, MeeusenSL, AinscowEK, and BrandMD. Suppressors of superoxide production from mitochondrial complex III. Nat Chem Biol, 11: 834–836, 2015.
44.
ParkHW, KimJY, ChoiSK, LeeYH, ZengW, KimKH, MuallemS, and LeeMG. Serine–threonine kinase with-no-lysine 4 (WNK4) controls blood pressure via transient receptor potential canonical 3 (TRPC3) in the vasculature. Proc Natl Acad Sci U S A, 108: 10750–10755, 2011.
45.
PonzoV, SoldatiL, and BoS. Resveratrol: a supplementation for men or for mice?. J Transl Med, 12: 158, 2014.
46.
PotenzaMA, MarasciuloFL, TarquinioM, TiravantiE, ColantuonoG, FedericiA, KimJA, QuonMJ, and MontagnaniM. EGCG, a green tea polyphenol, improves endothelial function and insulin sensitivity, reduces blood pressure, and protects against myocardial I/R injury in SHR. Am J Physiol Endocrinol Metab, 292: E1378–E1387, 2007.
47.
PrabhuSD and FrangogiannisNG. The biological basis for cardiac repair after myocardial infarction: from inflammation to fibrosis. Circ Res, 119: 91–112, 2016.
48.
RobichMP, OsipovRM, NezafatR, FengJ, ClementsRT, BianchiC, BoodhwaniM, CoadyMA, LahamRJ, and SellkeFW. Resveratrol improves myocardial perfusion in a swine model of hypercholesterolemia and chronic myocardial ischemia. Circulation, 122: S142–S149, 2010.
49.
RoseRA, HatanoN, OhyaS, ImaizumiY, and GilesWR. C-type natriuretic peptide activates a non-selective cation current in acutely isolated rat cardiac fibroblasts via natriuretic peptide C receptor-mediated signalling. J Physiol, 580: 255–274, 2007.
50.
Ruiz-VillalbaA, SimónAM, PogontkeC, CastilloMI, AbizandaG, PelachoB, Sánchez-DomínguezR, SegoviaJC, PrósperF, and Pérez-PomaresJM. Interacting resident epicardium-derived fibroblasts and recruited bone marrow cells form myocardial infarction scar. J Am Coll Cardiol, 65: 2057–2066, 2015.
51.
SahebkarA, SerbanC, UrsoniuS, WongND, MuntnerP, GrahamIM, MikhailidisDP, RizzoM, RyszJ, SperlingLS, LipGY, and BanachM;Lipid and Blood Pressure Meta-analysis Collaboration Group. Lack of efficacy of resveratrol on C-reactive protein and selected cardiovascular risk factors—results from a systematic review and meta-analysis of randomized controlled trials. Int J Cardiol, 189: 47–55, 2015.
52.
SalibaY, KaramR, SmayraV, AftimosG, AbramowitzJ, BirnbaumerL, and FarèsN. Evidence of a role for fibroblast transient receptor potential canonical 3 Ca2+ channel in renal fibrosis. J Am Soc Nephrol, 26: 1855–1876, 2015.
53.
SantiagoJJ, DangerfieldAL, RattanSG, BatheKL, CunningtonRH, RaizmanJE, BedoskyKM, FreedDH, KardamiE, and DixonIM. Cardiac fibroblast to myofibroblast differentiation in vivo and in vitro: expression of focal adhesion components in neonatal and adult rat ventricular myofibroblasts. Dev Dyn, 239: 1573–1584, 2010.
54.
ScalbertA and WilliamsonG. Dietary intake and bioavailability of polyphenols. J Nutr, 130: 2073S–2085S, 2000.
55.
SchleiferH, DoleschalB, LichteneggerM, OppenriederR, DerlerI, FrischaufI, GlasnovTN, KappeCO, RomaninC, and GroschnerK. Novel pyrazole compounds for pharmacological discrimination between receptor-operated and store-operated Ca(2+) entry pathways. Br J Pharmacol, 167: 1712–1722, 2012.
56.
SchulzE, WenzelP, MünzelT, and DaiberA. Mitochondrial redox signaling: interaction of mitochondrial reactive oxygen species with other sources of oxidative stress. Antioxid Redox Signal, 20: 308–324, 2014.
SeoK, RainerPP, Shalkey HahnV, LeeDI, JoSH, AndersenA, LiuT, XuX, WilletteRN, LeporeJJ, MarinoJPJr., BirnbaumerL, SchnackenbergCG, and KassDA. Combined TRPC3 and TRPC6 blockade by selective small-molecule or genetic deletion inhibits pathological cardiac hypertrophy. Proc Natl Acad Sci U S A, 111: 1551–1556, 2014.
59.
SinghA, HildebrandME, GarciaE, and SnutchTP. The transient receptor potential channel antagonist SKF96365 is a potent blocker of low-voltage-activated T-type calcium channels. Br J Pharmacol, 160: 1464–1475, 2010.
60.
SinghCK, LiuX, and AhmadN. Resveratrol, in its natural combination in whole grape, for health promotion and disease management. Ann N Y Acad Sci, 1348: 150–160, 2015.
61.
SpinaleFG and ZileMR. Integrating the myocardial matrix into heart failure recognition and management. Circ Res, 113: 725–738, 2013.
62.
ThandapillySJ, LouisXL, BehbahaniJ, MovahedA, YuL, FandrichR, ZhangS, KardamiE, AndersonHD, and NetticadanT. Reduced hemodynamic load aids low-dose resveratrol in reversing cardiovascular defects in hypertensive rats. Hypertens Res, 36: 866–872, 2013.
63.
ThandapillySJ, WojciechowskiP, BehbahaniJ, LouisXL, YuL, JuricD, KopilasMA, AndersonHD, and NetticadanT. Resveratrol prevents the development of pathological cardiac hypertrophy and contractile dysfunction in the SHR without lowering blood pressure. Am J Hypertens, 23: 192–196, 2010.
64.
TomidaT, HiroseK, TakizawaA, ShibasakiF, and IinoM. NFAT functions as a working memory of Ca2+ signals in decoding Ca2+ oscillation. EMBO J, 22: 3825–3832, 2003.
65.
TraversJG, KamalFA, RobbinsJ, YutzeyKE, and BlaxallBC. Cardiac fibrosis: the fibroblast awakens. Circ Res, 118: 1021–1040, 2016.
66.
Tresserra-RimbauA, RimmEB, Medina-RemónA, Martínez-GonzálezMA, López-SabaterMC, CovasMI, CorellaD, Salas-SalvadóJ, Gómez-GraciaE, LapetraJ, ArósF, FiolM, RosE, Serra-MajemL, PintóX, MuñozMA, GeaA, Ruiz-GutiérrezV, EstruchR, and Lamuela-RaventósRM;PREDIMED Study Investigators. Polyphenol intake and mortality risk: a re-analysis of the PREDIMED trial. BMC Med, 12: 77, 2014.
67.
van AmerongenMJ, Bou-GhariosG, PopaE, van ArkJ, PetersenAH, van DamGM, van LuynMJ, and HarmsenMC. Bone marrow-derived myofibroblasts contribute functionally to scar formation after myocardial infarction. J Pathol, 214: 377–386, 2008.
68.
VennekensR. Emerging concepts for the role of TRP channels in the cardiovascular system. J Physiol, 589: 1527–1534, 2011.
69.
VisioliF. The resveratrol fiasco. Pharmacol Res, 90: 87, 2014.
70.
WangB, XiongS, LinS, XiaW, LiQ, ZhaoZ, WeiX, LuZ, WeiX, GaoP, LiuD, and ZhuZ. Enhanced mitochondrial transient receptor potential channel, canonical type 3-mediated calcium handling in the vasculature from hypertensive rats. J Am Heart Assoc, 6: e005812, 2017.
71.
YueZ, XieJ, YuAS, StockJ, DuJ, and YueL. Role of TRP channels in the cardiovascular system. Am J Physiol Heart Circ Physiol, 308: H157–H182, 2015.
ZittC, StraussB, SchwarzEC, SpaethN, RastG, HatzelmannA, and HothM. Potent inhibition of Ca2+ release-activated Ca2+ channels and T-lymphocyte activation by the pyrazole derivative BTP2. J Biol Chem, 279: 12427–12437, 2004.
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